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Toru Shimazu

Publications and source records attributed to Toru Shimazu.

13 recordsLinked to original sources

Mutation induction in cultured human cells after low-dose and low-dose-rate gamma-ray irradiation: detection by LOH analysis.

To study the genetic effects of low-doses and low-dose-rate ionizing radiation (IR), human lymphoblastoid TK6 cells were exposed to 30 mGy of gamma-rays at a dose-rate of 1.2 mGy/hr. The frequency of early mutations (EMs) in the thymidine kinase (TK) gene locus was determined to be 1.7 x 10(-6), or 1.9-fold higher than the level seen in unirradated controls. These mutations were analyzed with a loss of heterozygosity (LOH) detection system, a methodology which has been shown to be sensitive to the effects of radiation. Among the 15 EMs observed after IR exposure, 8 were small interstitial-deletion events restricted to the TK gene locus. However, this specific type of event was not found in unirradiated controls. Although these results were observed under the limited conditions, they strongly suggest that the LOH detection system can be used for estimating the genetic effects of a low-dose IR exposure delivered at a low-dose-rate.

Cell Line↗

Short-term hypergravity does not affect protein-ubiquitination and proliferation in rat L6 myoblastic cells.

We previously reported that spaceflight (STS-90) and tail-suspension stimulated muscle protein ubiquitination and accumulated the degradation fragments. However, in space experiments the side-effects of hypergravity on samples are inevitable during the launch of a space shuttle into space or the reentry. To examine whether hypergravity also caused protein-ubiquitination in skeletal muscle cells, we exposed rat myoblastic L6 cells to various hypergravity conditions. Immunoblot analysis showed that the centrifugation at 2, 3, 30 or 100 G for 10 min did not increase the amount of ubiquitinated proteins in L6 cells, whereas the centrifugation at 100 G for 1 or 2 hrs significantly induced the protein-ubiquitination. In contrast, heat shock protein 70 (HSP70), another stress-responsive protein, in L6 cells was accumulated only by centrifugation at 100 G for more than 10 min. Short-term (10 min) hypergravity including 3 or 100 G did not affect the proliferation and morphological changes in L6 cells. Our present results suggest that the ubiquitination of muscle proteins is less sensitive to hypergravity than the induction of HSP70, and that the effect of hypergravity on protein-ubiquitination and proliferation of skeletal muscle cells may be negligible, as far as its duration is short-term.

Animals↗

Effects of running exercise during recovery from hindlimb unloading on soleus muscle fibers and their spinal motoneurons in rats.

The effects of hindlimb unloading and recovery with or without running exercise on morphological and metabolic properties of soleus muscle fibers and their spinal motoneurons in rats were investigated. Ten-week-old rats were hindlimb suspended for 2 weeks and thereafter were rehabilitated with or without voluntary running exercise for 2 weeks. A decreased percentage of type I fibers and atrophy of all types of fibers were observed after hindlimb unloading. In addition, decreased oxidative enzyme activity of all types of fibers was observed after hindlimb unloading. In contrast, an improvement in the decreased percentage of type I fibers, decreased fiber cross-sectional area, and decreased fiber oxidative enzyme activity was observed after recovery with running exercise, but not without running exercise. There were no changes in the number, cell body size, or oxidative enzyme activity of motoneurons innervating the soleus muscle after hindlimb unloading or recovery with or without running exercise. These results indicate that running exercise is beneficial for the recovery of the decreased percentage of type I fibers and the atrophy and decreased oxidative enzyme activity of all types of fibers in the soleus muscle induced by hindlimb unloading and that there are no changes in morphological or metabolic properties of spinal motoneurons innervating the soleus muscle following decreased or increased neuromuscular activity.

Animals↗

Animal habitats for space experiments.

There has been little opportunity for flight experiments using small animals, due to delay of construction of the International Space Station. Therefore, proposals using small animals have been unfortunately excepted from International Space Life Sciences Experiment application opportunity since 2001. Moreover, NASA has changed their development plan of animal habitats for space experiments according to changes of the U.S. space policy and the outlook is not so bright. However, international researchers have been strongly requesting the opportunity for space experiments using small animals. It will be also important for Japanese researchers to make a request for the opportunity. At the same time, researchers have to make an advance in ground based studies toward space experiments and to respond future application opportunities immediately. In this symposium, we explain the AEM (Animal Enclosure Module), the RAHF (Research Animal Holding Facility), and the AAH (Advanced Animal Habitat). It will be helpful for investigators to have wide knowledge of what space experiment is technically possible. In addition, the sample share program will be introduced into our communities. The program will provide many researchers with the organs and tissues from space-flown animals. We will explain the technical aspect of sample share program.

Animals↗

Development and verification of hardware for life science experiments in the Japanese Experiment Module "Kibo" on the International Space Station.

Japan Aerospace Exploration Agency (JAXA) has developed a cell biology experiment facility (CBEF) and a clean bench (CB) as a common hardware in which life science experiments in the Japanese Experiment Module (JEM known as "Kibo") of the International Space Station (ISS) can be performed. The CBEF, a CO2 incubator with a turntable that provides variable gravity levels, is the basic hardware required to carry out the biological experiments using microorganisms, cells, tissues, small animals, plants, etc. The CB provides a closed aseptic operation area for life science and biotechnology experiments in Kibo. A phase contrast and fluorescence microscope is installed inside CB. The biological experiment units (BEU) are designed to run individual experiments using the CBEF and the CB. A plant experiment unit (PEU) and two cell experiment units (CEU type1 and type2) for the BEU have been developed.

Animals↗

Growth-related changes in cell body size and succinate dehydrogenase activity of spinal motoneurons innervating the rat soleus muscle.

Cell body sizes and oxidative enzyme (succinate dehydrogenase) activities of spinal motoneurons innervating the soleus muscle were determined in rats ranging in postnatal age from 3 to 13 weeks. The soleus motoneurons were labeled by a retrograde neuronal tracer, nuclear yellow. The mean cell body sizes of motoneurons increased from 3 to 7 weeks of age, while the mean succinate dehydrogenase activities of motoneurons decreased from 3 to 7 weeks of age. There were no changes in mean cell body size or mean succinate dehydrogenase activity of motoneurons from 7 to 13 weeks of age. An inverse relationship between cell body size and succinate dehydrogenase activity of motoneurons was observed, irrespective of age. These results indicate that motoneurons innervating the rat soleus muscle show the adult pattern of cell body size and succinate dehydrogenase activity at an earlier stage of postnatal growth, 7 weeks of age.

Aging↗

Growth and development, and auxin polar transport of transgenic Arabidopsis under simulated microgravity conditions on a three-dimensional clinostat.

Growth and development, and auxin polar transport in Arabidopsis thaliana transformed with iaaH gene were studied under simulated microgravity conditions on a three-dimensional (3-D) clinostat. Simulated microgravity conditions on a 3-D clinostat did not affect the number of rosette leaves but promoted the growth and development (fresh weight of plant and the elongation of flower stalk) of transformants. Final growth of transformants under simulated microgravity conditions on a 3-D clinostat was almost equivalent to that grown on 1 g conditions in the presence of 1 micromoles IAM (indole-3-acetamide). The activities of auxin polar transport in the segments of flower stalk (inflorescence axis) of transformants grown on 1 g conditions were significantly promoted by the addition of IAM. Interestingly, simulated microgravity conditions on a 3-D clinostat also promoted the activities of auxin polar transport of transformants grown on the medium with or without IAM. Based on the results in this study, transgenic plants may not have an efficient homeostatic mechanism for the control of growth and development, and auxin polar transport activity in microgravity conditions in space.

Arabidopsis↗

Optical microscopy of Arabidopsis seedlings fixed in non-fresh FAA using Kennedy Fixation Tubes.

Samples will be kept in non-fresh fixative for longer period than usual in Kennedy Space Center Fixation Tubes (KFT) when they will be fixed for microscopy in the International Space Station (ISS). It is necessary to examine characteristics of samples prepared under such conditions and to improve preservation of the samples. In this study, morphology of Arabidopsis tissues prepared under such conditions was examined under a light microscope. Deformation of cell shape was observed in tissues with well-developed intercellular spaces when they were fixed in non-fresh fixative for longer period in KFTs.

Acetic Acid↗

Ground based verification of JAXA hardware for life science experiments.

Japanese new space agency (merger of NASDA, ISAS and NAL), JAXA (Japan Aerospace Exploration Agency) has been developing the Plant Experiment Unit (PEU) and the Cell Experiment Unit (CEU), which will be used within the Cell Biology Experiment Facility (CBEF) in "Kibo" Japanese Experiment Module (JEM) of ISS. They can also be operated within the Clean Bench (CB) in Kibo. We report the preliminary results of ground based verification experiments using the PEU and the CEU. Six units of PEU/CEU in microgravity section and 4 units of PEU/CEU in control-g section, will be operated within CBEF.

Cell Physiological Phenomena↗

Point of a space experiment proposal.

JAXA will solicit research proposals for space flight experiments that would be conducted for less than three years after the selection. In principle, available samples will be limited to Arabidopsis and C. elegans and flight hardware and protocol of space flight experiment will be pre-fixed. Proposals using different combinations of species and flight hardware will not be acceptable. Besides scientific issues, it is very important for proposer to write an impressive proposal. Hypothesis basis research proposal is the accepted standard. Reviewers will dislike a descriptive and unfocused research proposal without hypothesis. Ground preparation experiments, which are not related directly to space experiments, should not be included in the solicitation.

Animals↗

Effect of space flight on the frequency of micronuclei and expression of stress-responsive proteins in cultured mammalian cells.

Results of past space experiments suggest that the biological effect of space radiation could be enhanced under microgravity in some cases, especially in insects. To examine if such a synergistic effect of radiation and microgravity also exists in human cells, frequencies of chromosome instability and cellular levels of several stress-responsive proteins were analyzed in cultured human and rodent cells after space flight. Human (MCF7 and AT2KY), mouse (m5S) and hamster (SHE) cell lines were loaded on the Space Shuttle Discovery (STS-95 mission) and grown during a 9-day mission. After landing, the micronuclei resulting from abnormal nuclear division and accumulation of stress-responsive proteins such as p53 and mitogen-activated protein kinases (MAPKs), which are involved in radiation-induced signal transduction cascades, were analyzed. The frequencies of micronuclei in all the four mammalian cell strains tested were not significantly different between flight and ground control samples. Also, the cellular amounts of p53, p21 (WAF1/SDI1/CIP1) and activated (phosphorylated) forms of three distinct MAPKs in MCF7 and m5S cells of flight samples were similar to those of ground control samples. These results indicated that any effect of space radiation, microgravity, or combination of both were not detectable, at least under the present experimental conditions.

Animals↗